LOCUS       sequence-432668-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supression of shcircSMAD2(2,7)_2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Liu L, Neve M, Perlaza-Jimenez L, Xi X, Purcell J, Hawdon A, Conn 
            SJ, Zenker J, Tamayo P, Goodall GJ, Rosenbluh J
  TITLE     Systematic loss-of-function screens identify pathway-specific 
            functional circular RNAs.
  JOURNAL   Nat Cell Biol. 2024 Aug;26(8):1359-1372. doi: 
            10.1038/s41556-024-01467-y. Epub 2024 Aug 2.
  PUBMED    39095657
REFERENCE   2  (bases 1 to 7084)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41556-024-01467-y"; journalName: "Nat Cell Biol"; date: 
            "2024-08"; volume: "26"; issue: "8"; pages: "1359-1372"
COMMENT     Sequence Label: shRNA_circSMAD2_2
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(295..894)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(378..398)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     875..894
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(916..1426)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(979..997)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    1475..1592
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(1689..1937)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1747..1766)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1917..1937)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(2235..2279)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2464..2697
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    3190..3315
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3362..3542
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(3543..3769)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(3797..3817)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3797..3815)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3836..3852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3836..3852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3849..3871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3860..3876
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3884..3890,3891..3908,3909..3914))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3929..3950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4067..4084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4238..4826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4318..4337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4997..5788,5789..5857))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5620..5639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5858..5962)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5988..6443)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6125..6146)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6337..6356
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6570..6592
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6584..6601
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6585..6601
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6611..6630
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6611..6629
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      6650..6785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6825..6946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6836..6855)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6890..6909
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             join(7018..7084,1..167)
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt
       61 ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca
      121 agcaaaaagc agatcttgtc ttcgttggga gtgaattagc ccttccagtc cccccttttc
      181 ttttaaaaag tggctaagat ctacagctgc cttgtaagtc attggtctta aaggtaccga
      241 tgcatggggt cgtgcgctcc tttcggtcgg gcgctgcggg tcgtggggcg ggcgtcaggc
      301 accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt
      361 gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa
      421 ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag
      481 acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt
      541 gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct
      601 caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg
      661 cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag
      721 cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg
      781 atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc
      841 cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatggtaag
      901 ctccggtgga tccccctggg gagagaggtc ggtgattcgg tcaacgaggg agccgactgc
      961 cgacgtgcgc tccggaggct tgcagaatgc ggaacaccgc gcgggcagga acagggccca
     1021 cactaccgcc ccacaccccg cctcccgcac cgccccttcc cggccgctgc tctcggcgcg
     1081 ccctgctgag cagccgctat tggccacagc ccatcgcggt cggcgcgctg ccattgctcc
     1141 ctggcgctgt ccgtctgcga gggtactagt gagacgtgcg gcttccgttt gtcacgtccg
     1201 gcacgccgcg aaccgcaagg aaccttcccg acttaggggc ggagcaggaa gcgtcgccgg
     1261 ggggcccaca agggtagcgg cgaagatccg ggtgacgctg cgaacggacg tgaagaatgt
     1321 gcgagaccca gggtcggcgc cgctgcgttt cccggaacca cgcccagagc agccgcgtcc
     1381 ctgcgcaaac ccagggctgc cttggaaaag gcgcaacccc aaccccctcg agccgcggcc
     1441 aaagtggatc tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt
     1501 tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac
     1561 tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtctcg
     1621 aggtcgagaa ttcaaaaaac acaggttcga tacaagaggc tcgagcctct tgtatcgaac
     1681 ctgtgtccgg tgtttcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt
     1741 caagttacgg taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac
     1801 ccaagaaatt attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc
     1861 aaattaattc caattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc
     1921 atgggaaata ggccctcggt gaagggggcg gccgctcgag gctagtctcg tgatcgatac
     1981 cgtcgagatc cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct
     2041 tctattcctt cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat
     2101 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
     2161 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
     2221 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
     2281 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
     2341 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
     2401 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
     2461 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
     2521 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
     2581 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
     2641 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
     2701 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
     2761 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
     2821 tctcctcctc caggtctgaa gatcagcggc cgcttgctgt gcggtggtct tacttttgtt
     2881 ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg
     2941 cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg
     3001 aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc
     3061 aggattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata
     3121 ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg
     3181 cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt
     3241 tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt
     3301 cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca
     3361 ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca
     3421 cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc
     3481 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
     3541 ccgtttatgt atcgagctag gcacttaaat acaatatctc tgcaatgcgg caattcagtg
     3601 gttcgtccaa tccatgtcag acccgtctgt tgccttccta ataaggcacg atcgtaccac
     3661 cttacttcca ccaatcggca tgcacggtgc tttttctctc cttgtaaggc atgttgctaa
     3721 ctcatcgtta ccatgttgca agactacaag agtattgcat aagactacat taagcttgca
     3781 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     3841 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3901 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3961 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4021 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4081 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4141 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4201 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4261 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4441 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4561 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4621 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4681 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4741 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4801 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5161 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5221 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5281 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5341 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5401 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5461 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5521 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5581 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5641 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5701 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5761 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5821 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5881 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5941 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     6001 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     6061 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     6121 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     6181 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     6241 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     6301 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     6361 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     6421 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
     6481 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6541 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     6601 tgagcgcgcg taatacgact cactataggg cgaattgggt acgtccctag gcctccaaaa
     6661 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     6721 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     6781 gggatgggcg gagttagggg cgggatagct agagccagac atgataagat acattgatga
     6841 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     6901 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttcctc tcactctctg
     6961 atattcattt ctttgcaagt tataaatact gaataataag atgacatgaa ctactactgc
     7021 tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca
     7081 acag
//
